Development and evaluation of an IS711-based loop mediated isothermal amplification method (LAMP) for detection of Brucella spp. on clinical samples

Pérez Sancho, Marta; García-Seco Romero, María Teresa; Arrogante, L; García, N; García Benzaquén, Nerea; Martínez Alares, Irene; Díez Guerrier, Alberto Antoine; Perales, A; Goyache Goñi, Joaquín; Domínguez Rodríguez, Lucas José; Álvarez Sánchez, Julio

Development and evaluation of an IS711-based loop mediated isothermal amplification method (LAMP) for detection of Brucella spp. on clinical samples

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Development and evaluation of an IS711-based loop mediated isothermal amplification method (LAMP) for detection of Brucella spp on clinical samples.pdf (337.44 KB)

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https://www.sciencedirect.com/science/article/pii/S0034528813001811?via%3Dihub

Publication date

2013

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ELSEVIER SCI LTD

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URI

https://hdl.handle.net/20.500.14352/93391

Citation

Pérez-Sancho, M., García-Seco, T., Arrogante, L., García, N., Martínez, I., Diez-Guerrier, A., Perales, A., Goyache, J., Domínguez, L., & Alvarez, J. (2013). Development and evaluation of an IS711-based loop mediated isothermal amplification method (LAMP) for detection of Brucella spp. on clinical samples. Research in veterinary science, 95(2), 489–494. https://doi.org/10.1016/j.rvsc.2013.05.002

Abstract

DNA-based methods have emerged as an additional tool for Brucella infection-confirmation at a herd level. However, their implementation may require the use of specialized equipment. In this context the recently developed loop-mediated isothermal amplification (LAMP) technique may constitute an additional and cost-effective tool for rapid and specific DNA detection, especially in low income areas. In the present study the usefulness of a newly developed LAMP assay aiming at the multicopy-IS711 sequence was assessed on a variety of clinical samples (n = 81 from abortions and ewes; cattle, n = 3; swine, n = 4) that were analyzed in parallel using real-time PCR and bacteriology. Although overall sensitivities obtained with the three methods were comparable (p > 0.05), our results highlighted the complementarity between bacteriology and molecular-based methods for increased sensitivity. Significant differences (p < 0.05) were observed with all techniques depending on the nature of the sample. Our results demonstrate the potential of the IS711-LAMP technique for direct Brucella detection.

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2013 Elsevier Ltd. All rights reserved; Financiación: AGL-2009-12674 (Ministerio de Ciencia e Innovación). Consejería de Educación de la Comunidad de Madrid y Fondo Social Europeo. EU FP7 ANTIGONE Project Number 278976.